Department of Reproductive Medicine, Ghent University Hospital, De Pintelaan 185, 9000 Ghent, Belgium.
Reprod Biomed Online. 2010 Dec;21(6):769-75. doi: 10.1016/j.rbmo.2010.07.001. Epub 2010 Nov 3.
Parthenogenetic reconstruction is one major strategy to create patient-specific stem cells. The aim of this study was to find the best artificial activation protocol for parthenogenetic activation of mouse and human oocytes comparing different methods. In a first set of experiments, in-vivo matured mouse oocytes and human failed-fertilized, in-vitro and in-vivo matured oocytes were artificially activated by a chemical (ionomycin) or electrical stimulus. In a second set of experiments, a combination of activating agents (electrical pulses followed by ionomycin or SrCl(2)) was applied in an aim to improve developmental competence. All embryos were evaluated daily until day 6 after activation. Mouse blastocysts were differentially stained to evaluate blastocyst quality. For mouse oocytes and human failed-fertilized oocytes, blastocyst development was significantly higher after electrical activation (P<0.05). For human in-vitro and in-vivo matured oocytes, blastocyst formation was only obtained after electrical activation of in-vitro matured oocytes. After combining activating agents, no differences in development could be observed. In conclusion, this study revealed that for both mouse and human oocytes development to the blastocyst stage was significantly better after electrical activation compared with chemical activation. Combining activating agents had no further positive effect on developmental potential.
孤雌生殖重建是一种产生患者特异性干细胞的主要策略。本研究的目的是通过比较不同方法,找到用于小鼠和人卵母细胞孤雌激活的最佳人工激活方案。在一组实验中,体内成熟的小鼠卵母细胞和体外受精失败的人类卵母细胞、体内和体外成熟的卵母细胞分别通过化学(离子霉素)或电刺激进行人工激活。在第二组实验中,应用激活剂(电脉冲后加离子霉素或 SrCl(2))组合,以提高发育能力。所有胚胎均在激活后第 6 天进行每日评估。用不同的方法对小鼠囊胚进行染色,以评估囊胚质量。对于小鼠卵母细胞和人类体外受精失败的卵母细胞,电激活后囊胚发育显著提高(P<0.05)。对于人类体外和体内成熟的卵母细胞,仅在体外成熟的卵母细胞进行电激活后才获得囊胚形成。在组合激活剂后,发育没有观察到差异。总之,本研究表明,与化学激活相比,电激活对小鼠和人卵母细胞的发育至囊胚阶段有显著的促进作用。联合使用激活剂对发育潜能没有进一步的积极影响。